Purification and properties of DNA polymerase from Mycobacterium tuberculosis H37Rv.
DNA polymerase has been purified approximately 2000-fold from Mycobacterium tuberculosis H37Rv. The purified preparation was homogeneous by electrophoretic criteria and has a molecular weight of 135 000. The purified enzyme resembles Escherichia coli polymerase I in its properties, being insensitive to sulfhydryl drugs and possessing 5',3'-exonuclease activity in addition to polymerase and 3',5'-exonuclease activities. However, it differs from the latter in its sensitivity to higher salt concentration and DNA intercalating agents such as 8-aminoquinoline. The polymerase exhibited maximal activity between 37--42 degrees C and pH 8.8--9.5. The polymerase was stable for several months below 0 degree C. However, the 5',3'-exonuclease activity was more labile. The effects of different metal ions, polyamines and drugs on the polymerase activity are presented.
D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D007700:Kinetics; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D013379:Substrate Specificity
D003609:Dactinomycin; D004259:DNA-Directed DNA Polymerase